The use of mass spectrometry to examine the formation and hydrolysis of the phosphorylated form of phosphoglycerate mutase

Electrospray mass spectrometry has been used to study the formation and hydrolysis of the phopshorylated forms of two phosphoglycerate mutases, The half-life of the enzyme from Saccharomyces cerevisiae was 35 min at 20 degrees C in 10 mM ammonium bicarbonate, pH 8.0, Addition of 1 mM 2-phosphogly-collate reduced this value by at least 100-fold, The phosphorylated form of the enzyme from Schizosaccharomyces pombe was much less stable with a half-life of less than 1 min, The results are discussed in terms of the kinetic properties of the enzymes, Mass spectrometry would appear to be a powerful method to study the formation and breakdown of phosphorylated proteins, processes which are of widespread significance in regulatory mechanisms.